PMID:15169890

Citation	Correa, RG, Tergaonkar, V, Ng, JK, Dubova, I, Izpisua-Belmonte, JC and Verma, IM (2004) Characterization of NF-kappa B/I kappa B proteins in zebra fish and their involvement in notochord development. Mol. Cell. Biol. 24:5257-68
Abstract	Although largely involved in innate and adaptive immunity, NF-kappa B plays an important role in vertebrate development. In chicks, the inactivation of the NF-kappa B pathway induces functional alterations of the apical ectodermal ridge, which mediates limb outgrowth. In mice, the complete absence of NF-kappa B activity leads to prenatal death and neural tube defects. Here, we report the cloning and characterization of NF-kappa B/I kappa B proteins in zebra fish. Despite being ubiquitously expressed among the embryonic tissues, NF-kappa B/I kappa B members present distinct patterns of gene expression during the early zebra fish development. Biochemical assays indicate that zebra fish NF-kappa B proteins are able to bind consensus DNA-binding (kappa B) sites and inhibitory I kappa B alpha proteins from mammals. We show that zebra fish I kappa B alphas are degraded in a time-dependent manner after induction of transduced murine embryo fibroblasts (MEFs) and that these proteins are able to rescue NF-kappa B activity in I kappa B alpha(-/-) MEFs. Expression of a dominant-negative form of the murine I kappa B alpha (mI kappa B alpha M), which is able to block NF-kappa B in zebra fish cells, interferes with the notochord differentiation, generating no tail (ntl)-like embryos. This phenotype can be rescued by coinjection of the T-box gene ntl (Brachyury homologue), which is typically required for the formation of posterior mesoderm and axial development, suggesting that ntl lies downstream of NF-kappa B . We further show that ntl and Brachyury promoter regions contain functional kappa B sites and NF-kappa B can directly modulate ntl expression. Our study illustrates the conservation and compatibility of NF-kappa B/I kappa B proteins among vertebrates and the importance of NF-kappa B pathway in mesoderm formation during early embryogenesis.
Links	PubMed PMC419862 Online version:10.1128/MCB.24.12.5257-5268.2004
Keywords	Animals; Cells, Cultured; DNA, Complementary/genetics; Gene Expression Regulation, Developmental; Humans; I-kappa B Proteins/genetics; I-kappa B Proteins/metabolism; Mice; Mice, Knockout; NF-kappa B/genetics; NF-kappa B/metabolism; Notochord/embryology; Notochord/metabolism; Phenotype; Recombinant Proteins/genetics; Recombinant Proteins/metabolism; Species Specificity; T-Box Domain Proteins/genetics; T-Box Domain Proteins/metabolism; Zebrafish/embryology; Zebrafish/genetics; Zebrafish/metabolism; Zebrafish Proteins/genetics; Zebrafish Proteins/metabolism
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